Microbiocidal nitrogen derivatives of halogenated biphenyls

ABSTRACT

MICROBIOCIDAL BIS-AMINO SUBSTITUTED HALOGENATED BIPHENYLS, AS WELL AS THE NITROGEN DERIVATIVES THEREOF SUCH AS QUATERNARY AMMONIUM COMPOUNDS, AMINE OXIDES, IMIDAZOLINES, AMIDES, ENAMINES, AMPHOLYTES, AND THE LIKE.

United States Patent Oflice 3,733,421 MICROBIOCIDAL NITROGEN DERIVATIVES OF HALOGENATED BIPHENYLS John J. Merianos, Jersey City, Edward Griffin Shay, Belle Mead, Phillip Adams, Murray Hill, and Alfonso N. Petrocci, Glen Rock, N..l., assignors to Millmaster Onyx Corporation, New York, N.Y.

No Drawing. Continuation-impart of application Ser. No. 883,636, Dec. 9, 1969, now Patent No. 3,663,620. This application May 18, 1970, Ser. No. 38,513

Int. Cl. A01n 9/20 US. Cl. 424330 1 Claim ABSTRACT OF THE DISCLOSURE Microbiocidal bis-amino substituted halogenated biphenyls, as well as the nitrogen derivatives thereof such as quaternary ammonium compounds, amine oxides, irnidazolines, amides, enamines, ampholytes, and the like.

This is a continuation-inpart of co-pending application Ser. No. 883,636, filed Dec. 9, 1969, now US. Pat. No. 3,663,620.

This invention relates to bis-amino derivatives of halogenated biphenyls, and mixtures thereof with monoamino compounds, which have marked antimicrobial efficacy; and it particularly relates to the use of these compounds for treating industrial water; for the preservation of metal-working fluids; for use in germicidal detergenttype surface-active agents, including both synthetic and natural detergents, but especially soaps; and for the preservation of cosmetics and the like.

The compounds employed in the invention comprise the bis-amino substituted halogenated biphenyls having the general structure in which in and n are integers from to 4, X is a halogen, R and R are either hydrogen, alkyl or hydroxylalkyl, or form the residue of an amine or polyamine, or part of a cyclic amine structure, R and R may be either the same or different from each other and may be part of a polyamine; and there is at least one halogen atom; and with or without a content of unreacted halobiphenyl. In every case X may be fluorine, chlorine, bromine or iodine.

It was heretofore believed that only the monoaminosubstituted compounds of this type were satisfactory to obtain the microbiocidal results. However, it has now been found that the bis-compounds, by themselves are very effective for such purposes. In addition, mixtures of the his compounds with the mono compounds are not only also effective but are often more effective when the proportion of the his compound is larger than the mono compound.

In addition to their own inherent microbiocidal activity, these amino compounds can be used as intermediates for the preparation of a variety of other nitrogen compounds having antimicrobial powers. Among such are quaternary ammonium compounds, tertiary amine oxides, irnidazolines, ampholytes, Schiffs bases, amides, metal-amino coordination compounds, and, in general, antimicrobial derivatives of the amino group or groups to which the polyhalobiphenyl portion additionally contributes its own characteristics, such as, for example, a high degree of fireretardancy.

The bis compounds are prepared by reacting either ammonia or an amine with a halobiphenyl in such a manner that the mol ratio of ammonia or amine to halobiphenyl is at least :1 and preferably at least 10:1. On the other hand, when the mol ratio is less than 5: 1, more 3,733,421 Patented May 15, 1973 of the monoamino compound is obtained. The monoamino compounds have the structure:

wherein m is an integer from 0 to 5, n is an integer from 0 to 4, X is a halogen, and there is at least one halogen atom, R and R being either selected from the group consisting of hydrogen, alkyl, and hydroxyalkyl, or the residue of an amine or a polyamine, or are part of a cyclic amine structure; and wherein R and R may be either the same or different, and may be part of a polyamine.

Where chlorinated compounds are disclosed in the examples, the fluorinated, brominated, or iodinated compounds may be substituted in each case for the chlorinated compounds to obtain generally the same desired results.

As a source of raw materials, the Aroclors (polychlorinated polyphenyls-Monsanto) provided a wide range of polychlorinated biphenyls, e.g. Aroclor 1260 (said to contain 60% of chlorine) and Aroclor 1268 (said to contain 68% of chlorine). Others with other chlorine content, especially Aroclors 1232, 1242, 1248, 1254, and 1262, having chlorine contents of about the percent by weight represented by the last two digits in each case, have also been satisfactorily reacted, but the reaction rates of the members containing lesser amounts of chlorine are lower. In the case of these members, the addition of catalytic amounts of cuprous chloride or ferric chloride accelerate the reaction.

The halobiphenylamino compounds of the present invention may be employed either as the free amines or as their salts of inorganic or organic acids in order to improve their compatibility with aqueous, oily or solvent systems or to satisfy pH requirements. Such acids may include, for example, hydrochloric, sulfuric, phosphoric, acetic, lauric, oleic, gluconic, oxalic, tartaric, citric, benzoic and substituted benzoic, paratoluene sulfonic, sulfonic, sulfamic, and the like.

The following examples illustrate the preparation of both the monoamino and the bis-amino compounds. They are intended to be descriptive, but not necessarily limitative of the invention, except as claimed:

EXAMPLE 1 A one liter, three-necked round-bottomed flask, fitted with an agitator, a thermometer and a reflux condenser, was charged with 360 grams (or one mol) of Aroclor 1260, and 310 grams (of three mols) of diethylene triamine. The mixture was stirred thoroughly for 8 to 10 hours at 150l65 C. at atmospheric pressure until a snatch sample, taken under active agitation and dissolved in mixed isopropanol and water, indicated, by argentometric titration, that the reaction was essentially complete. The amine hydrochloride formed in the course of the reaction tended to separate to the top.

The reaction mixture was cooled to about 50 C. and it was then drowned in aqueous hydrochloric acid, the amount of the latter being adjusted to give a pH of about 5 to 6. A clear solution was obtained, with a concentration of 25% to 45% by weight at room temperature. A gummy mass separated, which liquified on heating to alkaline to about pH 10 by the addition of caustic soda. A gummy mass separated ,which liquified on heating to about 70 C. or higher. This was washed several times with hot water, and then dried.

The yield was essentially the theoretical, and calculated as the rnonosubstituted product, by equivalent weight titration in non-aqueous medium with perchloric acid, as crude N-(pentachlorobiphenyl)-diethylene triamine.

EXAMPLE 2 'In the apparatus of Example 1, 500 grams (or one mol) of Aroclor 1268 (assaying 70% Cl) and 310 grams (or three mols) of diethylene triamine were reacted in a similar manner by heating slowly to about 120 C., then at 125130 C. for about four hours until the reaction was complete, as indicated by argentometric titration. The same finishing procedure as in Example 1 was utilized to yield the theoretical amount of N-(nonachlorobiphenyl) diethylene triamine.

EXAMPLE 3 In apparatus similar to that of Example 1, and in a similar manner, 146 grams (or 0.5 mol) of Aroclor 1248 and 256 grams (or 2.5 mol) of diethylene triamine were reacted by heating at 180-190 C. for 36 hours. After 15 hours, reaction was 50% complete, and after 36 hours, it was 99% complete, as determined by argentometric titration. The excess amine was distilled off and 150 grams was recovered. The residue was treated with aqueous hydrochloric acid to yield a 30% active clear solution of amber color at pH 6.6.

The product so obtained was made alkaline with caustic soda, and the separated N-(trichlorobiphenyl)-diethylene triamine was washed and dried. It calculated 95%, by perchloric acid titration in non-aqueous medium EXAMPLE 4 In the same way, instead of being reacted with diethylene triamine, the polychlorobiphenyls were reacted with aliphatic and alicyclic polyamines such as ethylene diamine; 1,3-diaminopropane, dimethylaminopropylamine, triethylene tetramine, tetraethylene pentamine, hexamethylene-diamine, isophorone-diamine, et'hanolamine, diethanolamine, hydroxyethylethylene diamine, piperazine and aminoethyl piperazine, to yield the corresponding polychlorobiphenylamino derivatives, as the monosubstituted compounds. The reaction may also be carried out instead with primary or secondary alkylamines in similar fashion. Representative of these alkylamines are hexyl, octyl, nonyl, undecyl, dodecyl, tetradecyl, hexadecyl, octadecyl and octadecenyl amines.

The above amino derivatives of polychlorobiphenyls were employed as intermediates for the synthesis of nitrogen-bearing derivatives exemplified by the following types:

EXAMPLE 5 The products of Examples 1-4, such as do not already contain a tertiary amino group, were first alkylated on an amino nitrogen by methods well known to the art (for example, by reaction with formaldehyde and formic acid) to form tertiary amines; these were then quaternized, as in the present example; or converted to their tertiary amine oxides, as in the following Example 7.

The quaternizing agents included such compounds as the alkyl halides, the substituted or unsubstituted benzyl halides, a-chloromethylnaphthalene, dimethyl sulfate, and the like.

For example, 55 grams (or 0.1 mol) of N-(nonachlorobiphenyl) aminopropyl-N,N-dimethylamine and 17 grams (or'0.1 mol) of n-hexylbromide were dissolved in 100 grams of isopropanol. The mixture was heated under reflux at atmospheric pressure at 8595 C. for from 12 to 24 hours until reaction was essentially complete. The clear red to brown solution contained 87% of the theoretical of N-(nonachlorobiphenyl) aminopropyl-N,N-dimethyl-N-hexyl ammonium bromide.

Similarly, the n-octyl, n-decyl, n-undecyl, n-dodecyl, ntetradecyl, n-hexadecyl and n-octadecyl bromides were reacted to form the corresponding quaternary ammonium bromides.

Other quaternary ammonium salts were prepared, em-

4 ploying respectively, benzyl, ethylbenzyl, dichlorobenzyl and trichlorobenzyl halides; trimethylbenzyl halide; 2- methy-S-isopropyl benzyl halide; and chloromethylnaphthalene. In each instance, the halide may be chloride or bromide.

EXAMPLE 6 258 grams (or one mol) of Aroclor 1242 and 322 grams (or about 7.5 mols) of ethanol were charged into an autoclave. To this was added grams (or five mols) of liquid ammonia, after which the autoclave was sealed and heated with agitation to about 275 C. under autogenous pressure for about five hours. The autoclave was cooled and the excess ammonia was released into an absorber. The charge was drowned in dilute hydrochloric acid and the unreacted Aroclor was separated. The filtrate was made alkaline with caustic soda and the crude product was separated and dried, as a brown solid, in 131 grams or 55% yield of dichlorobiphenylamine. The nitrogen content was 5.6% (theory=5.85%).

EXAMPLE 7 From the tertiary amines prepared as in Example 5, the corresponding tertiary amine oxides were prepared. For example, 55 grams (or 0.1 mol) of N-(nonachlorobiphenyl)-aminopropyl-N,N-dimethylamine was dissolved in grams of isopropanol and the solution was heated at 6075 C. at atmospheric pressure under reflux and agitation, while adding gradually 20 grams (or 0.12 mol) of 35% aqueous hydrogen peroxide. The heating was continued for about 3 to 4 hours until the reaction was essentially complete and most of the excess hydrogen peroxide had decomposed. The product, N-(nonachlorobiphenyl) aminopropyl N',-N'-dimethylamine-N-oxide so obtained exhibited antimicrobial activity.

Instead of hydrogen peroxide, organic peroxides or ozone may be used.

EXAMPLE 8 Into a one liter round-bottomed flask fitted with an agitator, a thermometer, and a Dean and Stark water collection apparatus, were charged 52.4 grams (or 0.1 mol) of N-(nonachlorobiphenyl) ethylenediamine and 14 grams (or 0.1 mol) of p-hydroxybenzoic acid, along with 500 ml. of dry toluene. The mixture was agitated and heated until the mixture refluxed freely, and about 1.7 ml. of water was collected, during about 3% to 4 hours.

About 250 m1. of the toluene was stripped off, and the residue was cooled in an ice bath. The precipitated product was filtered and dried. A Kjeldahl determination showed a nitrogen content of 4.13% (theoretical 4.36%).

This product, N-(nonachlorobiphenyl)-aminoethyl-N'- p-hydroxybenzamide proved effective at 0.1% in the preservation from microbial decomposition of a wheat gluten paste.

Similarly, salicyclic acid, 2,4,5-trichlorobenzoic acid, and nicotinic acid were reacted with N-(nonachlorobiphenyl) ethylene diamine and N-(pentachlorobiphenyl) diethylene triamine, to produce their corresponding amides.

EXAMPLE 9 The polychlorobiphenylamino derivatives of Examples 1-4, such as have a terminal primary amino group, were reacted with substituted or unsubstituted aldehydes, and particularly aromatic aldehydes, to produce Schiffs bases. These displayed moderate antimicrobial activity.

EXAMPLE 10 Metal-amino coordination products were prepared by methods exemplified by the following procedure, from the products of Examples 1-4:

13.6 grams (or 0.1 mol) of anhydrous zinc chloride and 300 ml. of dry toluene were charged into a round bottomed flask fitted with an agitator, a thermometer, a reflux condenser and a dropping funnel. The mixture was heated at 40 C. while gradually adding a mixture of 157 grams (or 0.3 mol) of N-(nonachlorobiphenyl) ethylenediamine and 600 ml. of toluene. When the addition was complete, the mixture was heated for about two hours at the reflux temperature at atmospheric pressure.

After cooling to room temperature, the suspension was filtered ed, to yield tris-[IQ-(nonachlorobiphenyl) ethylene diamine] ZnCl This product was particularly effective in the perservation of paint films against fungal attack.

EXAMPLE 11 The related compounds were prepared by substituting for zinc chloride other salts such as BCl CuCl AlCl MnCl MgCl CoCl and the like, the amount of each varying with the molecular weight.

Using essentially the same procedure as in Example 10, their corresponding metal-amino products were made using the monoamino substituted polyhalobiphenyls synthesized as in Examples 14, the amount of each varying with its respective molecular weight.

EXAMPLE 12 The polybromobipheuyls may be prepared by brominating biphenyl in the presence of catalysts such as iodine, iron or aluminum, and along with agents such as sulfuric acid or chlorine, by methods known to the art. Such are described, for example, in British Pat. No. 934,970, and German Pats. Nos. 1,136,683 and 1,161,547.

The bromobiphenyls react with the amines and with ammonia in the same way as the chlorobiphenyls.

EXAMPLE 13 The Aroclors may be converted to fluoriuated biphenyls, with or without one or more residue halogen atoms, by fluorination followed by dehydrohalogenation, by which process fluorine addition products of the chlorobiphenyls are formed, which are then rendered aromatic again by the reaction of caustic alkali, removing hydrogen and chlorine. This procedure is described in V. Grakauskas Direct Liquid Phase Fluorination of Halogenated Aromatic Compounds," in the Journal of Organic Chemistry, Vol. 34, No. 10, pp. 2835-39 (October 1969).

The polyfiuorobiphenyls react with amino compounds in the same way as the Aroclors, to yield the corresponding fiuoroderivatives.

EXAMPLE 14 The polyiodinated biphenyls are also prepared by methods known to the art, for example, by reaction of Aroclors with KI.

These, too, react with amino compounds to yield the corresponding iodobiphenyl derivatives.

EXAMPLE 15 In the apparatus of Example 1, 326 grams (or one mol) of Aroclor 1254 and 1,030 grams (or tenmols) of diethylene triamine were charged. The mixture was heated under agitation at the reflux temperature, about 205 C., at atmospheric pressure for about 40 hours.

A snatch sample was titrated argentometrically and was found to contain 5.5% of ionic chlorine. Theory for (trichlorobiphenyl)-bis-diethylene triamine is 5.25%.

The excess diethylene triamine was distilled off under 10-20 mm. pressure; the cooled residue was drowned in water containing caustic soda. The product layer was separated, washed and dried by heating at 100-120 C. at 10-20 mm. pressure to a brown paste. Titration with acid indicated an equivalent Weight of 126, the theoretical being 115.

EXAMPLE 16 In a similar manner, the bis-aminosubstituted derivatives of diethylene triamine with Aroclors 1242, 1248, 1260, 1262, and 1268 were prepared.

EXAMPLE 17 The bis-aminosubstituted derivatives of the above Aroclors were prepared with ammonia and the amines of Examples 1 to 4 and 6, employing the techniques described therein.

EXAMPLE 18 For antimicrobiocidal evaluation of certain of the above compounds, the Standard Broth Inhibition test method was employed. Aliquots of the test materials were added to appropriate broth culture media contained in test tubes so that various concentrations of the test material in culture media were obtained. The tubes so prepared were inoculated with either 24 hour broth cultures of the test bacteria, or 14 day aqueous spore suspensions of the test fungi, or 7 day broth cultures of the algae. The inoculated tubes were incubated as follows: bacteria for 72 hours at 37 C.; fungi for 14 days at 28 C.; algae for 7 days at 25 C. Following the aforementioned incubation period, the tubes were examined for the presence or absence of macroscopic growth. The lowest concentration of test material in the broth which does not permit macroscopic growth is designated as the Minimum Inhibitory level.

In the following tables, these abbreviations of the designations of the organisms are employed: E.c.=Es cherichia coli; S.F.=Streprococcus faecalis; Ps.a.-=Pseudom'onas aeruginosa; A.n.=Aspergillus niger; P.e.=Penicilium expansum; C.p.=Chl0rella pyrenoidosa.

Certain representative quaternary ammonium salts of Example 5 were tested for static level of inhibition. These were the N-(nonachlorobiphenyl) aminopropyl-N',N'-dimethyl-N-alkyl ammonium bromides. The values are in parts per million of the quaternary Certain of the derivatives of tri-, pentaand nonachlorobiphenyl amino compounds of Examples 1-4, were tested bacteriologically for minimum inhibition levels as the free amines and their salts. For the sake of brevity they are referred to in the following Table 2 in terms respectively of C1 C1 and C1 with respect to the parent amine, ethylene diamine is referred to as E.D.; propylene diamine is referred to as P.D.; diethylene triamine is referred to as D.T.; triethylene tetramine is referred to as T.T.; dimethylaminopropylamine is referred to as DMAPA; isophorone diamine is referred to as ID; and hexamethylene diamine is referred to as H.D.

TABLE 2 Compound E.c. Ps.a. SJ. Am. P.e. 0.1)

1 E.D 10 250 10 10 50 1 Cl DMAPA 50 5 50 500 1 D.T 1 100 10 1, 000 1, 000 1 C15, D.T 5 50 10 C15, T.'1 10 100 10 C15, I.D 000 1,000 1, 000 1,000 1,000 750 Gig, LD 000 1, 000 1, 000 1, 000 1, 000 750 Ch, P. 10 100 100 1,000 000 50 Clo, E.D 10 100 100 1,000 1,000 10 C13, D.T 1 5 5 10 10 1 C1 H.D 10 500 50 50 10 10 C10, H.D 50 1, 000 50 500 1,000 50 EXAMPLE 20 N- (nonachlorobiphenyl) -aminopropyl-N',N'-dimethylamine was formulated as a 0.5% solution, by weight, in an acetone-aqueous soap solution. A stock soap solution TABLE 3 N-(nonachlorobiphenyl) aminopropyl-N,N-dimethylamine Soap 150/200 No soap 10/50 The effectiveness of the aminosubstituted halobiphenyls as soap bacteriostat is clearly shown above, and also in the following test. This is especially significant since it is notorious that most soap bacteriostats are not very active against gram negative bacteria.

EXAMPLE 21 Sterile soap solution was prepared at 2.5% soap solids;

propyl-N',N-dimethylamine was added at levels of 1% and 0.5% by weight, respectively, to a triethanolamine salt of Antara LM500 (a complex organic phosphate ester produced by General Aniline and Film Corp.) and to Mahogany Sulfonate (a petroleum sulfonate produced by Humble Oil and Refining Co.), both being examples of metal-working fluids. These were then diluted, one part of lubricant to 24 parts of water; the preservative was therefore present at 200 p.p.m. and 400 p.p.m. levels in the diluted fluid.

Sterile, wide mouth four ounce jars were charged with 100 ml. of the prepared dilutions. A similar dilution but not containing antimicrobial agent was charged to serve as a blank, for each of the metal-working fluids.

A series of 24 hour broth cultures of Escherichia coli, Pseudomonas aeruginosa, Bacillus species, Proteus species and Aerohacter aerogenes were pooled, diluted with sterile broth and inoculated into the 100 ml. samples to provide 1 to 10 10 bacteria cells per ml. of the samples.

At weekly intervals up to nine weeks, the samples were examined to determine the number of viable organisms present. At the five week point, each jar was re-inoculated as before, and the testing continued. The counts are given in the following table in which the count is to be multiplied by 10 N (nonachlorobiphenyl) aminopropyl-N,N'-dimethy1- amine was added to aliquots of this solution at 0.1 by weight, or 1,000 parts per million in the solution, and at a soap to product ratio of 25:1. For purposes of comparison, Phisohex (a proprietary product of Winthrop Laboratories, Division of Sterling Drug, Inc.) containing surface-active agents and 3% of 2,2'-methylene-bis(3,4,6- trichlorophenol) was diluted with distilled Water, also at the 1,000 ppm. level of bacteriostatic agent.

100 ml. aliquots of the respective solutions were transferred to Erlenmeyer flasks maintained at 25 C. and were inoculated with 1 ml. of an aqueous suspension of a 24 hour agar growth of Escherichia coli.

At intervals of three and five minutes after inoculation, aliquots of each of the solutions were plated into nutrient agar to determine the number of surviving bacteria. The number of survivors after five minutes was compared with the number initially present, from which figures the percent of organisms killed was computed. The following percentages are the average of replicate counts.

TABLE 4 Percent of Escherichia coli killed in 5 minutes Phisohex 94.3325 C1 DMAPA 99.3631

The present compounds are equally effective in synthetic detergents such as the fatty alcohol sulfates, and ether sulfates, and the alkylaryl sulfonates. The Maprofix WA series and Maprofix TLS" series (Onyx Chemical Co.) are representative of the first; Maprofix ES of the ether sulfates; and Ultrawet L60 (Arco Chem. Co.) is representative of the sulfates. Both soaps and detergents are effective in proportions of between about 20/1 and 100/1 of the soap or detergent to the amino-halobiphenyl compounds.

EXAMPLE 22 0.5% by weight of N (nonachlorobiphenyl)-amino- The term metal-working fluids, as used herein, are those discusses in Metalworking Lubricants by E. L. H. Bastian (McGraw-Hill Co., 1951, pp. 5-56). The present compounds have biocidal utility when intermixed in biocidally effective amounts with the fluids disclosed in this publication.

EXAMPLE 23 The efficacy of mixtures of the mono-aminosubstituted halobiphenyls of Examples 1 to 4 with the bis-amino-substituted halobiphenyls such as those of Examples 15-16 was compared by bacteriological assay. Among the combinations were the following monoand bisderivatives halobiphenyls of diethylene triaminefwith and without the presence of the Aroclor from which they were prepared, namely, Aroclors 1242, 1248, 1254, 1260, and 1268.

In each case, the following mixtures were prepared:

other derivatives exemplified above may be applied to the preservation of industrial water, both for use in cooling towers, air-conditioners, humidifiers and de-humidifiers and the like, and for use in process water, as for example in paper manufacture, to control the growth of microorganisms and to prevent slime formation, which would tend to clog heat exchangers, to destroy wood in the structure of towers and the like, and which would cause discoloration of paper pulp.

The products of this invention are also effective preser- TABLE 9 Mixtures from Aroclor 1254" percent Organism Mono Bis Aroclor E. P8 a SJ! SJ. Am. P.e. C42

EXAMPLE 24 The (trichlorobiphenyl) bis diethylene triamine of Example was reacted with a series of fatty acids to form bis-imidazolines.

An agitated, round-bottomed flask fitted with a thermometer and a takeoff condenser was charged with 230 grams (or 0.5 mol) of the amino compound, and 150 grams (or 1 mol) of caprylic acid. The mixture was heated for about four hours at 160 C. at atmospheric pressure, during which time the greater part of two mols of water were distilled oif. The heating was continued for another half hour while drawing a vacuum to about 25 mm. pressure. On cooling, the loss in weight was found to be about 34 grams, indicating the formation and removal of two mols of water per mol of fatty acid resulting from the amidification and the closing of the ring. After acetylating an aliquot of the product, titration with standard perchloric acid indicated essentially complete conversion to the imidazoline. The yield was essentially 100%, as his- [l-(trichlorobiphenyl aminoethyl)-2-heptyl] imidazoline in the form of a light-amber paste.

In the same manner, bis-imidazolines were prepared, substituting for caprylic acid a corresponding molar amount respectively, of acetic, propionic, butyric, valeric, caproic, oenanthic, pelargonic, capric, undecylenic, lauric, myristic, palmitic, and stearic acids.

EXAMPLE 25 The bis-imidazolines may be converted into their salts by treatment with the appropriate acid; or they may be quaternized if desired by reaction with agents such as are listed in Example 6.

The above bis-imidazolines exhibited distinct microbiocidal activity.

In general, unless otherwise specified, wherever parts or proportions have been used herein, it refers to parts by weight.

The compounds or mixtures thereof or their salts and vatives against microbial growth and action in cosmeitc preparations such as creams, lotions, shampoos, and the like, when present therein in a proportion of about 0.25% to 2% by weight of the composition, preferably about 0.5%, to prevent discoloration, putrefaction, phase separation, etc., and to prevent infection resulting from the use of such contaminated cosmetics.

They may also be introduced into metal-working fluids, to preserve them against microbial action which results in decomposition and putrefaction, or breaking of emulsions, or to prevent dermatitis resulting from contact with spoiled cutting and grinding oils and the like.

They may also be used in detergents, including bar soap, as for example, for a deodorant and germicidal soap.

They may also be applied to inanimate hard or soft surfaces, such as hospital walls, floors and the like and to textiles, rugs and the like, to render them bacteriostatic or bacteriocidal.

The invention claimed is:

1. A method for controlling bacteria and fungi which comprises applying to said bacteria and fungi a bacteriocidally and fungicidally effective amount of a bis-aminosubstituted halogenated biphenyl compound selected from the group consisting ofN-(trichlorobiphenyl)-diethylene triamine, N (pentachlorobiphenyl)-diethylene triamine and N-(nonachlorobiphenyl)-diethyleue triamine.

References Cited Chemical Abstracts, vol. 53 (1959), p. 5573b. Chemical Abstracts, vol. 58 (1963), p. 12388g-l2389. Chemical Abstracts, vol. 59 (1963), p. 15444h-15445.

ALBERT T. MEYERS, Primary Examiner V. D. TURNER, Assistant Examiner US. Cl. X.R. 

